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ID   YCP50      preliminary; circular DNA; SYN; 7950 BP.
XX
AC   X70276; K03077; ATCC37419;
XX
DT   01-JUL-1993 (Rel. 7, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   Saccharomyces/E.coli plasmid vector YCp50 - complete.
XX
KW   cloning vector.
XX
OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
XX
RN   [1]
RC   from YCp50 & yeast genes
RC   pRSa4-8 from YEp13 & yeast genes
RC   pCT80 from pUC12 & pRSa4-8, topoisomerase I gene
RA   Thrash C., Bankier A.T., Barrell B.G., Sternglanz R.;
RT   "Cloning, characterization, and sequence of the yeast DNA
RT   topoisomerase I gene";
RL   Proc. Natl. Acad. Sci. U.S.A. 82:4374-4378(1985).
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RN   [2]
RC   from YCp50
RA   Rose M., Novick P., Thomas J.H., Botstein D., Fink G.R.;
RT   "A Saccharomyces cerevisiae genomic plasmid bank based on a
RT   centromere-containing shuttle vector";
RL   Gene 60:237-243(1987).
XX
RN   [3]
RP   1-7950
RC   YCp50
RA   Stillman D.J.;
RT   ;
RL   Submitted (19-MAY-1993) by:
RL   Stillman D.J., University of Utah, Utah.
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RN   [4]
RC   GAL Sc4911 from GAL gene
RC   GAL Sc4812 from GAL gene
RC   pBM45 from YRp14 & GAL Sc4911 & GAL Sc4812
RC   galI-111, galI-112, galI-152, galI-227 from pBM45
RC   YCp50 from YCp19 & YIp5
RC   pBM125 from YCp50 & galI-152
RC   pBM150 from pBM125
RA   Johnston M., Davis R.W.;
RT   "Sequences that regulate the divergent GAL1-GAL10 promoter in
RT   Saccharomyces cerevisiae";
RL   Mol. Cell. Biol. 4:1440-1448(1984).
XX
CC   YCp50 was constructed by removing the EcoRI site in YCp19 between ARS1
CC   and CEN4 and cloning the PvuII/HindIII fragment into the PvuII site of
CC   YIp5.
CC   The PvuII site was not regenerated. [2]
CC   YC-type shuttle vector.
CC   The order of the major features in this plasmid is: HindIII - BamHI -
CC   SphI - URA3 - CEN4 - ARS1 - pMB1 ori - bla.
CC   Medium is 1227 LB plus ampicillin.
CC
CC   To pBR322, insert 1.1 kb HindIII-HindIII URA3 fragment into
CC   AvaI site with no overhangs and 10 G's added to nt 1428-1437
CC   and 10 C's added to nt 2601-2610 [dG/dC tailing] to make YIp5.
CC
CC   Ligate CEN4 from PvuII to EcoRI to ARS1 from EcoR1 to Hind III.
CC   Delete 190 nt around EcoRI site [85 nt from each side]. BalI site is
CC   deleted; perhaps it is intact in [2].
CC
CC   Insert CEN4-ARS1 fragment [PvuII - CEN4 - dead EcoRI - ARS1 -
CC   HindIII/klenowed] into PvuII site of YIp5. Change CAGCTG to CAGGTG
CC   so PvuII site is not regenerated and restriction site numbering
CC   of [2] is retained.
CC   NCBI gi: 406851
CC   NM (YCp50)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP (ATCC)
CC   HO (E.coli HB101)(Saccharomyces cerevisiae)(E.coli)
CC   CP ()
CC   FN (cloning)(shuttle)
CC   SE ()
CC   PA (pBR322)(YIp5)
CC   BR ()
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. YCp19 EcoRI 10600bp, between ARS1 and CEN4
FT                   S1 nuclease
FT                   fill in
FT                   PvuII-HindIII 2400bp, ARS1/CEN4/YRp17
FT                   \ YRp17 1494..-..4711
FT                   :Klenow
FT                   2. YIp5 PvuII 5541bp 3250..3250
FT                   -> YCp50 7950bp [no PvuII]"
FT   misc_feature    join(1..1429,2611..3247,5657..7950)
FT                   /note="from pBR322"
FT   misc_feature    join(1430..2600,3248..5656)
FT                   /note="from Saccharomyces cerevisiae"
FT   misc_feature    1..1429
FT                   /note="from pBR322; EcoRI to AvaII, blunted"
FT   misc_feature    1429..1438
FT                   /note="poly dG-dC tail"
FT   CDS             1439..2600
FT                   /note="ANT yeast URA3 gene"
FT   misc_feature    2601..2610
FT                   /note="poly dG-dC tail"
FT   misc_feature    2611..3247
FT                   /note="from pBR322; AvaII, blunted to PvuII;
FT                   PvuII site not regenerated"
FT   misc_feature    3248..4911
FT                   /note="yeast centromere CEN4"
FT   rep_origin      4912..5656
FT                   /note="ORI yeast ARS1"
FT   misc_feature    5657..7950
FT                   /note="from pBR322; PvuII to EcoRI"
FT   misc_binding    0..0
FT                   /note="SIT unique EcoRI-BglII-KpnI-XhoI-HpaI-SmaI-
FT                   ClaI-BstXI-NsiI(AvaIII)-
FT                   BamHI-HindIII-ApaI-NcoI-SalI-PvuI-NruI-SphI-XmaIII-
FT                   AsuII-BspmII-SpeI-AatII"
FT   rep_origin      0..0
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             0..0
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
FT   CDS             0..0
FT                   /note="ANT E. coli tetracycline resistance gene (tet)"
XX
SQ   Sequence 7950 BP; 2142 A; 1852 C; 1811 G; 2145 T; 0 other;
     gaattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt
     aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct
     cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac tgccgggcct
     cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct
     atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg
     ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc
     gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgacc
     gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
     cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcactgg
     accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
     gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatcgg ggggggggtt
     tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc tttgaaattt
     ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg
     atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     ttaattatat cagttattac ccgggaatct cggtcgtaat gatttttata atgacgaaaa
     aaaaaaaatt ggaaagaaaa cccccccccc gcagcgttgg gtcctggcca cgggtgcgca
     tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
     agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
     cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
     agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
     ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
     gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
     ccatgaacag aaattccccc ttacacggag gcatcaagtg accaaacagg aaaaaaccgc
     ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     ttaccgcagg tgggccattc tcatgaagaa tatcttgaat ttattgtcat attactagtt
     ggtgtggaag tccatatatc ggtgatcaat atagtggttg acatgctggc tagtcaacat
     tgagcctttt gatcatgcaa atatattacg gtattttaca atcaaatatc aaacttaact
     attgacttta taacttattt aggtggtaac attcttataa aaaagaaaaa aattactgca
     aaacagtact agcttttaac ttgtatccta ggttatctat gctgtctcac catagagaat
     attacctatt tcagaatgta tgtccatgat tcgccgggta aatacatata atacacaaat
     ctggcttaat aaagtctata atatatctca taaagaagtg ctaaattggc tagtgctata
     tatttttaag aaaatttctt ttgactaagt ccatatcgac tttgtaaaag ttcactttag
     catacatata ttacacgagc cagaaattgt aacttttgcc taaaatcaca aattgcaaaa
     tttaattgct tgcaaaaggt cacatgctta taatcaactt ttttaaaaat ttaaaatact
     tttttatttt ttatttttaa acataaatga aataatttat ttattgttta tgattaccga
     aacataaaac ctgctcaaga aaaagaaact gttttgtcct tggaaaaaaa gcactaccta
     ggagcggcca aaatgccgag gctttcatag cttaaactct ttacagaaaa taggcattat
     agatcagttc gagttttctt attcttcctt ccggttttat cgtcacagtt ttacagtaaa
     taagtatcac ctcttagagt taactatgag ataagcaagt atcatctcat ttcatttacc
     tgaagtcgag taaacagaaa atccaattgt tgatgaacct caatgactta gaactatcta
     tcggcagatc atataaagag gatttaggta cctagaggac tgtacctgga gtatatatat
     atatatatat atattatctc aactatagtc catagaggtt tctttcttga ggccttaaac
     tgctaaagaa tgatattggt ggaatgcaag caccaatctc tcttctttcg taactgttca
     tatacttcaa accaagaatg taacgggcat tgacccatcc aaaaccttca gtagctgccc
     ctttaaagtc agcaccttga ttaccgtatt ctgcttcaac acgatgagga tctgttcctc
     ttgtgacatc atatttttca accacaatac cattataatc gacaaaagcc tttgtcatca
     tgaaaagcca tctataagct agcctattcg ttacagttaa ataaccataa gaacggaggc
     cttcccaagc aagaatttga tggggtgccc aaccaaatgg atagtcccat tgtctaattg
     gtctcgaaat agaaattggg cctcgagaac gctccgtaca tgcagctaaa cctccaagca
     tctctaactt gggtagtgct ttctccacca ttttctgtgc ttgctccttc gtggcaagtc
     cagcccataa tgcccagaat gtagttgcgg attcgtatga cgttctgtgc ttgatttttg
     tgttgtagtc aaagaaaaac cccgactcgt catcccacat atatttggta attgatgagg
     caacgctaat tatcaacata tagattgtta tctatctgca tgaacacgaa atctttactt
     gacgacttga ggctgatggt gtttatgcaa agaaaccact gtgtttaata tgtgtcactg
     tttgatatta ctgtcagcgt agaagataat agtaaaagcg gttaataagt gtatttgaga
     taagtgtgat aaagttttta cagcgaaaag acgataaata caagaaaatg attacgagga
     tacggagaga ggtatgtaca tgtgtattta tatactaagc tgccggcggt tgtttgcaag
     accgagaaaa ggctagcaag aatcgggtca ttgtagcgta tgcgcctgtg aacattctct
     tcaacaagtt tgattccatt gcggtgaaat ggtaaaagtc aaccccctgc gatgtatatt
     ttcctgtaca atcaatcaaa aagccaaatg atttagcatt atctttacat cttgttattt
     tacagatttt atgtttagat cttttatgct tgcttttcaa aaggcctgca ggcaagtgca
     caaacaatac ttaaataaat actactcagt aataacctat ttcttagcat ttttgacgaa
     atttgctatt ttgttagagt cttttacacc atttgtctcc acacctccgc ttacatcaac
     accaataacg ccatttaatc taagcgcatc accaacattt tctggcgtca gtccaccagc
     taacataaaa tgtaagctct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     aggcgtatca cgaggccctt tcgtcttcaa
//